Puresyn, Inc. has developed purification processes for supercoiled plasmid DNA vaccine and gone therapy vectors utilizing its proprietary chromatographic resin, PolyFIo(r). In Phase I, a tandem chromatography processes was developed: hydrophobic interaction and PolyFIo. The process resulted in product of high purity and recovery that exceeds other chromatographic processes. We demonstrated this process removes significant amounts of host contaminants as well as other plasmid isoforms. The resulting product has improved biological activity as assessed in in vitro transfection assays. [unreadable] [unreadable] We are seeking Phase II funding to concentrate on supercoiled plasmid production to include large-scale purification up 5 grams per run without compromising purity, yield or biological activity. We will apply newly developed analytical methodologies to investigate the factors that may influence biological activity. Finally, we will conduct an in vivo mouse study to determine the correlation between purity and biological activity through the evaluation of immune and gone therapeutic responses to several vectors. [unreadable] [unreadable] Results of this research will have immediate impact for those engaged in the use of supercoiled vectors for DNA vaccines and gone delivery because it will allow the large-scale development of plasmid vectors. It will have positive commercialization implications for PolyFIo and Puresyn, Inc. [unreadable] [unreadable]